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Paths of Protein Trafficking Nuclear Protein Transport Mitochondrial & Chloroplast Transport Experimental Systems Overview of the Cytomembrane System The Endoplasmic Reticulum The Golgi Apparatus Vesicular Transport between Compartments Exocytosis Endocytosis and Lysosomes
Ribosomes in Cytosol
Completed protein via Posttranslational Import Goes to: Cytosol, Nucleus, Mitochondria, Plastids, Peroxisomes
Ribosomes with partially synthesized proteins: Attach to Endoplasmic Reticulum Protein is imported into ER via Cotranslational Import Protein is modified in ER and goes to:
Golgi Apparatus via Vesicular Transport Protein is further modified and goes to:
Secretory Vesicles
Vesicles from Golgi fuse with Early Endosomes to form Late Endosomes
Plasma Membrane
Experimental Systems
1. 2. 3. 4. Autoradiography, Pulse-Chase, and Green Fluorescent Protein Experiments Differential Staining Cell Fractionation Genetic Mutant Analysis
Mitochondrial/Chloroplast ProteinTransport
Mitochondrial proteins that are encoded by nuclear genes possess mitochondrial import sequences Transport from the cytosol into the mitochondria is mediated by a TOM complex and two TIM complexes The SAM complex mediates proper folding of outer membrane proteins with -barrel structures Another complex (OXA) mediates insertion of mitochondrial-encoded proteins into inner membrane & also contributes to insertion of some nuclear-encoded proteins Proteins must be unfolded, either by chaperones or by specialized unfolding proteins
Mitochondrial/Chloroplast ProteinTransport
ATP hydrolysis & a H+ gradient drive protein transport into mitochondria via a ratcheting mechanism Integral proteins possess stop transport sequences that interrupt the transport process to create the transmembrane domains; formation of the transmembrane domain may be either mediated by TIM23, TIM22 (specialized for multipass proteins), or OXA
Mitochondrial/Chloroplast ProteinTransport
Transport of chloroplast proteins is similar to mitochondrial proteins Thylakoid proteins require an extra thylakoid transport sequence & import proteins
Endoplasmic Reticulum
1. Functions of the Smooth ER
Steroid Hormone Synthesis Detoxification in Liver Release of glucose from liver Sequestering calcium ions
Endoplasmic Reticulum
2. Functions of the Rough ER
Cotranslational import of proteins destined for the ER-Golgi pathway Synthesis of membrane lipids and generation of lipid compositional asymmetry Protein glycosylation: Synthesis of the core portion of an N-linked oligosaccharide
Golgi Apparatus
Modification of N-linked oligosaccharides Synthesis of O-linked oligosaccharides Phosphorylation of mannose (on N-linked oligosaccharide) on proteins targeted for lysosomes Sorting of proteins into secretory vesicles or primary lysosomes
Exocytosis
Secretory vesicles (from the trans-Golgi) are targeted to the plasma membrane, with which they fuse. The soluble contents of the vesicles are released to the outside, and the vesicle membrane becomes part of the PM.